CN1205474C - Biosensor - Google Patents

Biosensor Download PDF

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CN1205474C
CN1205474C CNB028018796A CN02801879A CN1205474C CN 1205474 C CN1205474 C CN 1205474C CN B028018796 A CNB028018796 A CN B028018796A CN 02801879 A CN02801879 A CN 02801879A CN 1205474 C CN1205474 C CN 1205474C
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electrode
counter electrode
sample
working electrode
substrate
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CN1463361A (en
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谷池优子
池田信
吉冈俊彦
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松下电器产业株式会社
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/001Enzyme electrodes
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/001Enzyme electrodes
    • C12Q1/005Enzyme electrodes involving specific analytes or enzymes
    • C12Q1/006Enzyme electrodes involving specific analytes or enzymes for glucose
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/26Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving oxidoreductase
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N27/00Investigating or analysing materials by the use of electric, electro-chemical, or magnetic means
    • G01N27/26Investigating or analysing materials by the use of electric, electro-chemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
    • G01N27/28Electrolytic cell components
    • G01N27/30Electrodes, e.g. test electrodes; Half-cells
    • G01N27/327Biochemical electrodes electrical and mechanical details of in vitro measurements
    • G01N27/3271Amperometric enzyme electrodes for analytes in body fluids, e.g. glucose in blood
    • G01N27/3272Test elements therefor, i.e. disposable laminated substrates with electrodes, reagent and channels

Abstract

本发明是提供一种即使是在测定极微量的试样量时,也能得到良好应答的高灵敏度生物传感器。 The present invention is to provide a measurement even when an extremely small amount of the amount of the sample, a favorable response can be obtained with high sensitivity biosensor. 本发明的生物传感器具有分支成多个的作用极和分支成多个的第1对极、将各分支片交替排列的第1绝缘性基板,具有第2对极、配置于和第1绝缘性基板相对位置上的第2绝缘性基板,含有氧化还原酶的试剂系以及形成在第1和第2绝缘性基板之间的试样供给通路,在供给通路内交替排列的作用极和第1对极的分支片、第2对极和试剂系外露。 The biosensor of the present invention has a first counter electrode, the first insulating substrate and the working electrode branches into a plurality of branched into a plurality of sheets are alternately arranged in the respective branch, having a second counter electrode, and arranged on the first insulating a second substrate opposed to the position of the insulating substrate, the reagent system comprising an oxidoreductase and a sample supply pathway formed between the first and second insulating substrate and alternately arranged in the supply passage and the first pair of the working electrode branch pole piece, and a second electrode exposed to the reagent system.

Description

生物传感器 biological sensor

技术领域 FIELD

本发明涉及迅速且高精度地定量试样中所含基质的生物传感器。 The present invention relates to quickly and precisely quantify biosensor contained in the sample matrix.

背景技术 Background technique

作为蔗糖、葡萄糖等糖类定量分析法,已开发出用旋光度计法、比色法、还原滴定法和各种色谱法等方法。 Sucrose, glucose, sugars such quantitative analysis method, a method has been developed using a spin polarimetry, colorimetry, reduction titration and a variety of chromatography. 但是,无论对哪样糖类,这些方法特异性不太高,因此精度较差。 However, regardless of Which sugars, these methods are not too specific, and therefore less accurate. 如果用这些方法中的施光度计法,操作是简单,但是,操作时,受温度影响大,所以,施光度计法不合适通常作为人们在家庭等地方简单定量糖类的方法。 If these methods are applied photometry, the operation is simple, but, during operation, affected by temperature, therefore, generally administered polarimetry inappropriate places such as people in the family of simple sugars quantitative method.

近年来,利用酶具有的特异性催化作用的各种类型的生物传感器被开发。 Recently, using an enzyme having the catalytic specificity of the various types of biosensors have been developed.

以下,就作为试样中基质定量法一例的葡萄糖定量法加以说明。 Hereinafter, it will be described as an example of a method for quantifying glucose in a sample matrix of quantitative method. 一般大家熟知的是作为电化学的葡萄糖定量法是使用酶的葡萄糖氧化酶(EC 1.1.3.4:以下省略为GOD)和氧电极或是过氧化氢电极的方法(例如,铃木周一编的《バィォセンサ一(生物传感器)》講談社出版)。 Generally well known as an electrochemical glucose quantification is the use of the enzyme glucose oxidase (EC 1.1.3.4: hereinafter abbreviated to GOD) and an oxygen electrode or a method of hydrogen peroxide electrode (e.g., Shuichi Suzuki series "ba ィ ォ ã nn sa a (biological sensor) "published by Kodansha).

GOD把氧作为电子传递介质将基质β-D-葡萄糖选择性地氧化成D-葡糖酸-δ-内酯。 GOD oxygen to the substrate β-D- glucose selective oxidation -δ- D- gluconic acid lactone as an electron transfer medium. 在氧存在的情况下,氧化反应过程中,氧通过GOD被还原成过氧化氢。 In the presence of oxygen, the oxidation reaction process, oxygen is reduced to hydrogen peroxide by GOD. 通过氧电极来计测此氧的减少量,或者是通过过氧化氢电极来计测过氧化氢的增加量。 Measured by the oxygen electrode to reduce the amount of this oxygen, or by the hydrogen peroxide electrode measured by increasing the amount of hydrogen peroxide. 因为氧的减少量和过氧化氢的增加量与试样中的葡萄糖含有量成比例,所以可以从氧的减少量或过氧化氢的增加量来定量葡萄糖。 Because of the reduced amount of oxygen and increase the amount of hydrogen peroxide in the sample is proportional to the glucose content, it can be, or by increasing the amount of hydrogen peroxide from a reduced amount of oxygen to glucose quantification.

在上述方法中,利用酶反应特异性,可以高精度地定量试样中的葡萄糖。 In the above method, using an enzyme-specific reaction, it can be accurately quantify glucose in the sample. 但是上述方法有测定结果会受到试样所含氧的浓度的影响的缺点,在试样里没有氧存在的情况下,就不能进行测定,这也可以从其反应过程推测出来。 However, these methods have the measurement result is affected by the concentration of oxygen-containing specimen drawbacks without the presence of oxygen, can not be measured in the sample, it may be inferred from the reaction process.

为此,开发了不用氧作为电子传递介质,而用铁氰化钾、二茂铁衍生物、苯醌衍生物等有机化合物和金属络合物作为电子传递介质的新型葡萄糖传感器。 For this reason, without the development of oxygen as an electron transfer medium, and an organic compound and a metal complex with potassium ferricyanide, ferrocene derivatives, quinone derivatives, etc. as electron transfer glucose sensor of new medium. 这种类型的传感器将作为酶反应的结果而生成的电子传递介质的还原体在作用极上氧化,并可由此氧化电流量求出试样中所含葡萄糖浓度。 Reduced form electron transfer mediator sensors of this type as a result of enzyme reaction generated in the oxidation of the working electrode, and thereby determine the concentration of glucose oxidation current contained in the sample. 这时,在对极上进行电子传递介质氧化体的还原,进行生成电子传递介质还原体的反应。 In this case, reduction of oxidized electron transfer medium on the counter electrode, the electron transfer reaction medium for generating a reduced form. 通过用上述的有机化合物和金属络和物来代替氧作为电子传递介质,就可以在稳定状态下将已知量GOD和这些电子传递介质正确地加载在电极上,形成试剂层,并可以不受试样中氧浓度影响,高精度地定量葡萄糖。 Using the above organic compound and a metal complex and may be substituted by oxygen as an electron transfer medium, it is possible in a steady state known amount of GOD and the electron transporting media loaded correctly on the electrode, the reagent layer is formed, and is free to Effect of concentration of oxygen in the sample, glucose quantification with high accuracy. 这种情况下,由于还能够使含有氧和电子传递介质的试剂层以接近干燥的状态下与电极系一体化,所以,基于该技术的一次性葡萄糖传感器近年来受到众多关注。 In this case, since the reagent layer can also contain oxygen and an electron transfer mediator close to the electrode system is integrated with a dry state, so that a disposable glucose sensor based on this technology in recent years been a lot of attention. 其代表例是日本特许公报第2517153号中公开的生物传感器。 Representative examples of biosensors which Japanese Patent Publication No. 2517153 disclosed. 使用一次性葡萄糖传感器,只需往与测定器连接并可拆卸的传感器中注入试样,就能够很容易地用测定器测出葡萄糖浓度。 Using a disposable glucose sensor, and the measuring unit is connected only to the detachable sensor and injection of the sample, it can be easily detected by measuring the glucose concentration.

通过使用了上述葡萄糖传感器的测定方法,就可以容易地求出微升数量级试样里的基质浓度。 By using the above-described method for measuring a glucose sensor, it can be easily determined in a sample of the order of [mu] l substrate concentration. 但是,近年来,各方面期望着开发出一种能够测定出比1微升更少的极微量试样的生物传感器。 Recently, however, desirable aspects of the development of a biosensor less than 1 microliter sample of a very small amount can be measured. 现有的电化学生物传感器,在测定极微量试样时,因为试样里的葡萄糖量变得极微量,所以出现测定结果的灵敏度下降的情况。 Conventional electrochemical biosensor, when a very small amount in the measurement sample, since the amount of glucose in the sample becomes extremely small amount, where a decrease in sensitivity of the assay results so there.

因此,就开发出在基板上配置有2个由分支片交替排列而构成的多个分支的近似梳子形电极的生物传感器。 Thus, the development of approximately comb-shaped electrode on the biosensor substrate are arranged alternately arranged over two branches by the branch constituted by the sheet. 图7显示此生物传感器的电极系附近的剖面图。 Figure 7 shows a cross-sectional view of the vicinity of the electrode system of the biosensor. 该类型的生物传感器,在配置于基板5的第1电极1上,经氧化而生成的电子传递介质的氧化体就可能在邻接的第2电极3上被还原而恢复成还原体,此还原体还可能再次在邻接的第1电极1上被氧化。 This type of biosensor, on the first electrode disposed on the substrate 1 to 5, oxidized generated electron mediator in oxidized medium may be reduced and restored to the reduced form on the adjacent second electrode 3, this reduced form It may also be re-oxidized at the first electrode adjacent to 1. 因此,通过流动在第1电极1上的电流值上升来看,此传感器可以以比现有生物传感器好的灵敏度来定量葡萄糖。 Accordingly, the current value increases in the first electrode 1 through the flow point of view, the sensor sensitivity may be better than the prior quantified glucose biosensor.

这样的技术,不仅仅局限于定量葡萄糖,还可能应用在试样里含有其它基质的定量上。 Such techniques, is not limited to glucose quantitation and may also be used in quantitative sample contains other substrates.

但是近年来,为了在测定时能求出更微量化的必要试样量,各方面期望着有一种更高灵敏度的生物传感器。 But in recent years, in order to obtain the necessary amount of energy measurement even smaller amount of sample, aspects desirable to have a higher sensitivity of the biosensor.

因此,本发明目的是提供一种即使是在测定极微量的试样量时,也能得到良好应答的高灵敏度生物传感器。 Accordingly, an object of the present invention to provide a measurement even when an extremely small amount of the amount of the sample, a favorable response can be obtained with high sensitivity biosensor.

发明内容 SUMMARY

本发明的生物传感器具有分支成多个的作用极和分支成多个的第1对极、将各分支片交替排列的第1绝缘性基板,具有第2对极、配置于和第1绝缘性基板相对位置上的第2绝缘性基板,含有氧化还原酶的试剂系以及形成在第1和第2绝缘性基板之间的试样供给通路,在供给通路内交替排列的作用极和第1对极的分支片、第2对极和试剂系外露。 The biosensor of the present invention has a first counter electrode, the first insulating substrate and the working electrode branches into a plurality of branched into a plurality of sheets are alternately arranged in the respective branch, having a second counter electrode, and arranged on the first insulating a second substrate opposed to the position of the insulating substrate, the reagent system comprising an oxidoreductase and a sample supply pathway formed between the first and second insulating substrate and alternately arranged in the supply passage and the first pair of the working electrode branch pole piece, and a second electrode exposed to the reagent system.

在前述试样供给通路内,第2对极最好配置在与作用极相对的位置上。 In the sample supply pathway, the second counter electrode is preferably disposed at a position opposite to the working electrode.

本发明提供一种具有分支成多个的第1作用极和分支成多个的第1对极、将各分支片交替排列的第1绝缘性基板,具有分支成多个的第2作用极和分支成多个的第2对极、将各分支片交替排列的第2绝缘性基板,含有氧化还原酶的试剂系以及形成在第1和第2绝缘基板之间的试样供给通路的生物传感器。 The present invention provides a first working electrode and having a plurality of branches branched into a plurality of first counter electrode, the sheets are alternately arranged in each branch of the first insulating substrate, having a plurality of branches of the second working electrode and branched into a plurality of the second counter electrode, the second insulating substrate pieces alternately arranged in each branch containing an oxidoreductase reagent system and a sample supply pathway of the biosensor is formed between the first and second insulating substrate . 在所述试样供给通路内交替排列的第1作用极和第1对极的分支片、交替排列的第2作用极和第2对极的分支片以及试剂系外露。 A first electrode and a counter electrode branch piece, the role of the second electrode are alternately arranged and exposed on the second pole piece and a reagent branch lines are alternately arranged in the first action of the sample supply passage.

最好是第2对极配置在和第1作用极相对的位置上,且第2作用极配置在和第1对极相对的位置上。 Preferably the second position and a counter electrode disposed opposing action on the first electrode and the second working electrode disposed in the first position and the opposed counter electrode.

附图说明 BRIEF DESCRIPTION

图1是本发明葡萄糖传感器一实施方式的除去试剂层的分解立体图。 FIG 1 is an exploded perspective view of a glucose sensor was removed reagent layer according to the present invention, an embodiment of FIG.

图2是显示同一传感器试样供给通路内的电极排列的剖面图。 FIG 2 is a cross-sectional view of the electrode in the sample supply pathway of the same sensor arrangement.

图3是显示传感器试样供给通路内的电极排列的另一例的剖面图。 FIG 3 is a cross-sectional view of another embodiment of the electrode arrangement of the sensor in the sample supply path.

图4是本发明生物传感器另一实施方式的除去试剂层的分解立体图。 FIG 4 is an exploded perspective view of the biosensor reagent layer removed according to another embodiment of the present embodiment.

图5是显示同一传感器试样供给通路内的电极排列的剖面图。 FIG 5 is a cross-sectional view of the electrode in the sample supply pathway of the same sensor arrangement.

图6是本发明生物传感器又一实施方式的除去试剂层的分解立体图。 FIG 6 is an exploded perspective view of the biosensor reagent layer removed according to the present invention still further embodiment.

图7是显示现有生物传感器电极排列的剖面图。 FIG. 7 is a sectional view of the conventional biosensor electrode arrangement.

图8是显示本发明一实施方式的安装有传感器的测定装置的电路结构方块图。 FIG 8 is a graph showing the installation embodiment of the present invention is a circuit block diagram showing the measuring device sensors.

图9是显示本发明另一实施方式的安装有传感器的测定装置的电路结构方块图。 9 is mounted to another embodiment of the present invention is a circuit block diagram showing the measuring device sensors.

实施本发明的最佳方式以下参照附图就本发明生物传感器的实施方式加以说明。 BEST MODE FOR CARRYING OUT THE INVENTION The following embodiments will be described with reference to the drawings embodiment of the present invention is a biosensor.

第1基板和第2基板等的形状结构,电极的形状和材质,分支片的数目不受以下所示实施方式的限制。 Shaped structure of the first substrate and the second substrate or the like, the number of shape and material of the electrode, the branch piece is not limited the following embodiments illustrated embodiment.

实施方式1图1是本实施方式葡萄糖传感器的除去试剂层和界面活性剂层的纵向剖面图。 Embodiment 1 Figure 1 is a longitudinal sectional view of the reagent layer is removed and the surfactant layer glucose sensor of the present embodiment.

10表示由电绝缘性材料构成的第1基板。 10 denotes a first substrate made of electrically insulating material. 在此基板10上,通过光刻工艺,形成了由分支成多个的近似梳子形的作用极11及其导线12,和分支成多个的近似梳子形的第1对极13及其导线14构成的电极系。 On this substrate 10 by a photolithography process, the formation of approximately comb-shaped first counter electrode 13 and the wire, and branched into a plurality of approximated by a plurality of comb-shaped branches of the working electrode 11 and its lead 1214 electrode system configuration. 具体方法是:例如将钯溅镀到基板上,用保护膜覆盖此钯膜,接着,加上和电极系相同形状的掩膜,经曝光,显像后,将钯膜蚀刻,最后,除去保护膜,就形成设定形状的电极系。 The specific method is: for example, palladium sputtering onto the substrate, the palladium membrane covering this with a protective film, and then, with the same shape as the electrode system, and a mask, the exposed, after development, the palladium film is etched, and finally, removing the protective film is formed on the electrode system with a set shape. 在图中,作用极11和第1对极各以6片分支片表示,但不限于此。 In the drawings, a first working electrode and the counter electrode 11 to each of the branch piece 6 represented, but is not limited thereto. 如后述实施例所示,作用极11和第1对极也可由数10片分支片构成。 As shown in Examples described later, the role of the first electrode 11 and the counter electrode 10 may also be the number of branches constituting the sheet. 在由电绝缘性材料构成的第2基板30上,将钯溅射到此基板上而形成第2对极33及其导线34。 On the second substrate 30 made of electrically insulating material, palladium is sputtered on this substrate to form a counter electrode 33 and the second wire 34. 第2基板30有空气孔35。 Air holes 30 of the second substrate 35. 在第1基板10上,为了将该装置的端部与第2对极的导线34接触,开有导通孔17,而为了使装置的端部与作用极的导线12和第1对极13的导线14相连接,在第2基板30上,开有导通孔36和37。 On the first substrate 10, to the end of the contact device and the second counter electrode lead 34, vias 17 are opened, and in order that the end portion of the action means 12 and the electrode wires of the first counter electrode 13 the wire 14 is connected to the second substrate 30, via holes 36 are opened and 37.

由绝缘材料构成的隔板20上具有形成后述的试样供给通路的切口21。 A separator made of an insulating material having a notched supply passage described later is formed of 21 20. 将此隔板20粘贴固定在第1基板10上后,从切口21滴下试剂层形成液在电极系上,经干燥形成试剂层,此试剂层包含有氧化还原酶GOD和电子传递介质铁氰化钾。 This separator 20 is attached is fixed to the rear of the first substrate 10, a slit 21 is dropped from the liquid reagent layer on the electrode system, and dried to form a reagent layer, the reagent layer comprises an oxidoreductase and an electron transfer mediator GOD ferricyanide potassium. 在试剂层上,最好形成含有界面活性剂卵磷脂的界面活性剂层。 On the reagent layer, preferably forming surfactant layer comprises a surfactant lecithin.

接着,按照图1中点划线所示的位置关系将结合有隔板20的第1基板10与第2基板30相连接而组装成葡萄糖传感器。 Next, FIG. 1 according to the midpoint of the positional relationship shown in chain line in conjunction with a separator 10 of the first substrate 20 and second substrate 30 is assembled is connected to the glucose sensor. 然后,在第1基板和第2基板之间,在隔板20的切口21部分,形成试样供给通路。 Then, between the first substrate and the second substrate, the cutout portion 21 of the separator 20 to form a sample supply pathway. 此试样供给通路以切口21的开放端部23为试样供给口,以第2基板30的空气孔35为终端部。 This sample supply passage to the open end of the cutout portion 21 of the sample supply port 23 to the air hole of the second substrate 35 of the terminal portion 30.

在此试样供给通路中,电极系和第2对极配置在相对的位置上,而且,用隔板作用极11、第1对极13和第2对极33面对试样供给通路的面积(电极面积)由隔板20确定。 In the sample supply pathway, the electrode system and a second counter electrode disposed in opposite positions, and with the working electrode separator 11, a first electrode 13 and second electrode 33 facing the sample supply pathway area pairs (electrode area) is determined by the partition 20.

以下参照图8,说明用此传感器测定葡萄糖的测定装置。 Referring now to Figure 8, a measurement device using this sensor for measuring glucose.

图8左侧所示为上述传感器70。 As shown in FIG. 8 is a left side 70 of the sensor. 在图中,仅示出作用极导线12、第1对极导线14和第2对极导线34。 In the drawings, showing only the working electrode lead 12, a first counter electrode lead wires 14 and 34 the second pole pair. 另一方面,测定装置71具有和上述导线12、14和34分别相连接的连接器72、74和84。 On the other hand, the measuring apparatus 71 having a connector and the wires 12, 14 and 34 are connected to 72, 74 and 84. 连接器84经开关76与连接器74相连接,通过开关75而使连接器84、74与基准电位发生电路77相连接。 The connector 84 is connected to switch 76 via connection 74, switching circuit 77 occurs through the connector 84,74 coupled to the reference potential 75. 连接器72与电位发生电路82和电流/电压变换电路78相连接。 78 connector 72 is connected to the potential circuit 82 and a current / voltage conversion circuit occurs. 电流/电压变换电路78以连接在基准电位发生电路77上的对极为基准,施加正电位在作用极上时,将在作用极和对极之间通过电流变换成电压输出。 A current / voltage conversion circuit 78 is connected to the reference potential generating extremely reference, a positive potential is applied to the electrode 77 acts on the circuit, between the electrode and the working electrode currents into voltage output by the pair. 用A/D变换电路79将此输出电压变换成脉冲,而CPU80根据由A/D变换电路79输出的脉冲,算出试样中的基质含量,此计算值用LCD81表示。 Conversion A / D converting the output voltage of this circuit 79 into a pulse, and pulse circuit 79 according to the CPU80 outputs from the A / D conversion, the matrix content in the samples is calculated, this calculated value is expressed LCD81.

按上述将传感器70配置在测定装置71上,关闭测定装置开关76,使第1对极13与第2对极33短路,并同时关闭开关75。 The above-described sensor 70 disposed on the measuring device 71, switch off the measuring device 76, so that the short-circuit electrode 13 electrode 33 first pair and the second pair, and at the same time closes the switch 75. 如将含葡萄糖的试样与传感器端部的试样供给口23接触,由于毛细管现象,试样会很容易地到达试样供给通路内的试剂层。 As will sample sample supply port to the transducer portion 23 contacts the glucose-containing, capillary action, a sample can easily reach the reagent layer in the sample supply pathway. 如检测出试样到达了电极系,测定装置就启动,计时器开始计时。 The detected sample reaches the electrode system, the measuring apparatus is started, the timer is started. 试剂层一溶解于试样里,葡萄糖就被GOD氧化,与此同时,电子传递介质铁氰化钾被还原成亚铁氰化钾。 A reagent layer is dissolved in the sample, the GOD-glucose was oxidized, at the same time, electron transfer mediator potassium ferricyanide is reduced to potassium ferrocyanide. 在装置开始启动以后经过适当的时间,以对极为基准,电压300mv由电位发生电路82施加在作用极11上,在作用极11和对极之间会有将亚铁氰化钾氧化的电流通过。 After an appropriate time, in order to extremely reference voltage is applied to 300mv 82 acting on the electrode 11 by the potential generating circuit means after started, the working electrode 11 and the counter electrode will have a current between the oxidation by potassium ferrocyanide . 通过测定装置的电流/电压变换电路78的以下的运作,基于上述电流值用LCD81表示出葡萄糖浓度。 By following the operating current / voltage conversion circuit 78 of the measuring device, based on the current value of the glucose concentration was expressed LCD81.

图2所示为在本实施方式生物传感器的电极系近旁将电子传递介质氧化的电流的流动状态。 Figure 2 shows the vicinity of the electrode system in the biosensor according to the present embodiment, the electron transfer medium flowing current state of oxidation. 在本实施方式中,作用极11和第1对极13多个地分支,这些分支片交替排列,形成电极系,在此电极系的相对方向,配置有第2对极33。 In the present embodiment, the first working electrode 11 and the electrode 13 to a plurality of branches, the branch sheets are alternately arranged to form the electrode system in the direction opposite this electrode system, the second counter electrode is disposed 33. 通过这样的构成,在配置于第1基板10上的作用极11上被氧化而生成的电子传递介质的氧化体于邻接的第1对极13上被还原,与此同时,在与作用极11垂直的方向上扩散的电子传递介质的还原体也在配置于第2基板30上的第2对极33上被还原,恢复成还原体。 With this configuration, acting on the first substrate 10 disposed on the electrode 11 is oxidized to form the oxidized electron transfer medium to the first pair of adjacent upper electrode 13 is reduced, at the same time, the working electrode 11 with reduced form of the electron transfer mediator diffusion are arranged in a direction perpendicular to the second electrode 33 is reduced to a pair of the second substrate 30, restored to the reduced form. 另外,由于作用极上的扩散层生长被抑制,第2对极33上的氧化还原种的浓度就被反映在传感器的应答上。 Further, since the diffusion layer on the working electrode growth is inhibited, the second redox species concentration at the electrode 33 was reflected in the response of the sensor. 由于上述这些原因,和现有生物传感器相比较,本实施方式生物传感器的应答提高了。 For these reasons, as compared to existing biosensors, the present embodiment improves the response of the biosensor.

这里,第2对极最好只配置在与作用极相对的位置上,即,如图3所示,将第2对极33修整,形成具有多个分支片33a的梳子状。 Here, only the second counter electrode is preferably disposed at a position opposite to the working electrode, i.e., FIG. 3, the counter electrode 33 of the second trimming, forming comb-like sheet having a plurality of branches 33a. 在试样供给通路中,第2对极的分支片33a放置在与作用极分支片相对方向上。 In the sample supply passage, the second counter electrode branch pieces 33a of the working electrode is placed on the branch plate opposite directions. 由于这样的话,作用极正上方的第2对极近旁的电流密度变得更高等,可以认为在第2对极近旁的还原型电子传递介质的浓度会增高。 Since this is the case, immediately above the working electrode of the second electrode becomes higher in the vicinity of the current density and the like, that can transfer medium concentration in the second electron reduction vicinity of the electrode will be increased. 因为传感器的应答是依存于还原型电子传递介质浓度的,所以作为结果,就可以高灵敏度地定量基质。 Since the sensor response is dependent on the concentration of the reduced electron transfer mediator, so as a result, it can be quantified with high sensitivity matrix.

实施方式2图4是本实施方式葡萄糖传感器除去了试剂层和界面活性剂层的分解立体图。 Embodiment 2 FIG 4 is a glucose sensor according to the present embodiment is removed exploded perspective view of the reagent layer and surfactant layer.

以和实施方式1相同的顺序,在第1基板40上形成由分支成多个的近似梳子形的第1作用极41、第1作用极导线42、分支成多个的近似梳子形第1对极43和其导线44构成的第1电极系。 In the same order and manner as Embodiment 1, is formed by the branch into a plurality of approximately comb-shaped first working electrode 41, first working electrode lead 42 is branched into a plurality of approximately comb-shaped first pair on the first substrate 40 43 and its lead electrode 44 composed of a first electrode system. 在第2基板60上形成由分支成多个的近似梳子形的第2作用极61、第2作用极导线62、分支成多个的第2对极63和其导线64构成的第2电极系。 Is formed on the second substrate 60 by the second electrode 61 is branched into a plurality of action of approximately comb-shaped second working electrode lead 62, the second electrode system is branched into the second counter electrode lead 63 and 64 which constitute a plurality of . 如同实施方式1,作用极和对极的分支片数不受图示数所限。 As in Embodiment 1, the number of branches of the working electrode and the counter electrode plate is not limited by the number shown. 将空气孔65形成在第2基板60上。 The air hole 65 is formed on the second substrate 60. 为了将装置的端部和第2对极的导线62和第2作用极的导线64相连接,在第1基板上形成导通孔48和49,同样,为了使装置端部和第1作用极的导线42和第1对极的导线44相连接,于第2基板60上开了导通孔68和69。 To the end of the device and the second counter electrode lead wire 62 and the second working electrode lead 64 is connected, is formed on the first substrate vias 48 and 49, similarly, in order that the end of the device and the first working electrode the wires 42 and the first counter electrode lead 44 is connected to the opening of the vias 68 and 69 on the second substrate 60.

接着,在第1基板40上,贴加隔板50后,形成试剂层,按照图4中所示的点划线位置关系将第2基板60连接起来制成葡萄糖传感器。 Subsequently, on the first substrate 40, and diaphragm 50 attached, form a reagent layer, in accordance with the positional relationship shown in dot-dash line in FIG. 4 connects the second substrate is made of a glucose sensor 60. 隔板50上具有形成试样供给通路的切口51,其切口51的开放端部52成为试样供给口。 Having a slit 51 is formed a sample supply pathway of the separator 50, its open end 52 of the cutout 51 becomes sample supply port.

如上述制作,通过隔板50的切口51,在第1基板40和第2基板60之间形成试样供给通路,然后,如图4所示,在试样供给通路内第2对极63配置在第1作用极41的相对位置上,而第2作用极61配置在第1对极43的相对位置上。 Produced as described above, the cutout 51 through the separator 50, a sample supply pathway formed between the first substrate 40 and the second substrate 60, and then, as shown in the sample supply passage 63 of the second four-pole configuration the relative position of the first working electrode 41 on the second working electrode 61 is disposed at the first relative position of the counter electrode 43. 通过隔板50的切口51确定第1作用极41、第1对极43、第2作用极61和第2对极63面对试样供给通路的面积(电极面积)。 The first separator is determined by the action of the cutout 51 50 41 pole, a first counter electrode 43, second working electrode 61 and counter electrode 63 facing the second area of ​​the sample supply pathway (electrode area). 以和实施方式1的作用极11同等电极面积来形成本实施方式的传感器的第1作用极41和第2作用极61的合计电极面积。 In the working electrode 11 and the electrode area the same embodiment of the sensor according to the present embodiment is formed of a first embodiment of the working electrode 41 and second working electrode 61 of the total electrode area. 但是,由于在第2基板上配置有第2作用极61,和实施方式1传感器相比较,就形成更高密度的电极系,因此,和实施方式1相比,可以缩小切口51的大小,使试样量减少。 However, since the second substrate is disposed on the second working electrode 61, and compared to Embodiment 1 sensor, the electrode system is formed of a higher density, and therefore, compared to Embodiment 1 and Embodiment, the size of the cutout 51 can be reduced, so that reduce the amount of sample.

这里,第2对极最好配置在第1作用极的相对位置上,并且第2作用极也最好配置在第1对极的相对位置上。 Here, the second counter electrode is preferably disposed at a position opposite poles on the first action, and the second working electrode is also preferably arranged at opposite positions on a first counter electrode.

图5所示为本实施方式的生物传感器试样供给通路中的电极排列。 Biosensor electrode arrangement shown in the sample supply pathway of the embodiment of the present embodiment in FIG 5. 将交替排列每个配置在第1基板40上的第1作用极41和第1对极43,和每个配置于第2基板60上的第2作用极61和第2对极63,并且,第1作用极41和第2对极63处于相对方向上,第1对极43和第2作用极61也处于相对方向上。 The first action are alternately arranged each arranged on the first substrate 40 and the first electrode 41 counter electrode 43, and each arranged in the second action on the second substrate 60 and the second electrode 61 counter electrode 63, and, the first working electrode 41 and the second counter electrode 63 in the opposing direction of the first counter electrode 43 and second working electrode 61 also in the opposite direction. 因此,和图2所示生物传感器相比,在总作用极面积相同的情况下,就有可能更密地配置电极系。 Therefore, as compared to the biosensor shown in FIG. 2, at the same emitter area of ​​action of the total cases, there may be more densely arranged electrode system. 由此,因为可以减少试样供给通路体积,所以就能减少检测体的试样量。 Accordingly, since the sample supply pathway can be reduced volume, the amount of the sample can be reduced in the sample.

实施方式3图6是本实施方式葡萄糖传感器除去了试剂层和界面活性剂层的分解立体图。 Embodiment 3 FIG 6 is a glucose sensor according to the present embodiment is removed exploded perspective view of the reagent layer and surfactant layer.

本实施方式和实施方式1不同之处在于在第1基板10上形成参比极15和其导线16,以及为了使装置2个端部各自和作用极导线12和参比极15的导线16相连接,在第2基板30上形成导通孔38,除此之外,其余结构都和实施方式1相同。 The present embodiment and Embodiment 1 except that 16 relative to the reference electrode 15 and its lead 16, and to enable means two end portions each and a working electrode lead wire 12 and reference electrode 15 is formed on the first substrate 10 connected, via holes 38 are formed on the second substrate 30, except the same manner as embodiment 1 and the rest of the structure.

下面参考图9就用此传感器来测定葡萄糖的测定装置加以说明。 Referring to FIG. 9 using this glucose sensor measuring apparatus will be described.

在图9左侧示出了上述传感器80。 In the left side of FIG. 9 shows the sensor 80. 图中仅示出作用极导线12、第1参比极导线16、对极导线14和第2对极导线34。 Figure shows only the working electrode lead 12, a first ratio of the reference electrode lead 16, electrode wires 14 and to the second counter electrode lead 34. 另一方面,测定装置81具有分别和上述导线12、16、14和34相连接的连接器72、96、74和84,其中连接器74和84与电流发生电路97相连接;电位发生电路82和电流/电压变换电路78与连接器72相连接。 On the other hand, the measuring apparatus 81 has a connector 12, 16, respectively, and said conductors are connected 72,96,74 and 34 and 84, 74 and 84 wherein the current generating circuit connector 97 is connected; potential generating circuit 82 and a current / voltage conversion circuit 78 and the connector 72 is connected. 电流/电压变换电路78、A/D变换电路79和CPU80如同实施方式1中说明的测定装置进行相同运作。 A current / voltage conversion circuit 78, A / D conversion circuit 79 and CPU80 measuring apparatus as in Embodiment 1 will be explained in the same operation.

按上述将传感器80装配在测定装置81上,如将含有葡萄糖试样与传感器端部试样供给23相接触,由于毛细管现象,试样会容易地到达试样供给通路内的试剂层。 The above-described sensor 80 mounted on the measuring device 81, such as a sample in a sample containing glucose is supplied to the sensor contact end 23, due to the capillary phenomenon, the sample will easily reach the reagent layer in the sample supply pathway. 如检测到试样到达了电极系,测定装置启动,计时器就开始计时。 The detected sample reaches the electrode system, the measurement device is activated, a timer starts counting. 在装置开始启动以后经过适当的时间,以参比极15为基准,300mv电压由电位发生电路82施加在作用极11上,在作用极11和对极之间就有将亚铁氰化钾氧化的电流通过。 After the device started after appropriate time, the reference electrode 15 to reference, at 300mV applied voltage acts on the electrode 82 by the potential generating circuit 11, the working electrode 11 and there between the potassium ferrocyanide oxidation electrode the current through. 和实施方式1相同,通过测定装置的电流/电压变换电路78以下的运作,用LCD81表示出基于上述电流值的葡萄糖浓度。 And the same manner as Embodiment 1, by the operation of the current / voltage conversion circuit 78 of the following measurement apparatus, based on the current value by glucose concentration LCD81 shown.

以和实施方式1相同的原因,本实施方式生物传感器和现有的生物传感器相比,其应答值提高了。 In the same manner as Embodiment 1 and the reason, the present embodiment and the conventional biosensor biosensor compared to the response value increased. 另外,与不配置参比极的传感器相比,由于设置了参比极15,稳定了作用极11的电位,因此,就可能更高精度地测定。 Further, as compared with the reference sensor is not disposed over the electrode, since the potential of the reference electrode 15, working electrode 11 is stabilized, and therefore, it is possible to determine with higher accuracy.

本发明,作为第1基板和第2基板,只要用具有电绝缘性、保存和测定时具有足够刚性的材料即可。 The present invention, as the first substrate and the second substrate, provided with an electrically insulating, a material having sufficient rigidity during storage and measurement. 例如可以用聚乙烯、聚苯乙烯、聚氯乙烯、聚酰胺、饱和聚酯树脂等热塑性树脂,或者是尿素树脂、三聚氰胺树脂、酚醛树脂、环氧树脂、不饱和聚酯树脂等热固性树脂。 For example, polyethylene, polystyrene, polyvinyl chloride, polyamide, saturated polyester resin, thermoplastic resin, or urea resin, melamine resin, phenol resin, epoxy resin, unsaturated polyester resin, a thermosetting resin. 从和电极密合性角度来看,最好用聚对苯二甲酸乙二酯。 And an electrode from the viewpoint of adhesiveness, preferably with poly ethylene terephthalate. 隔板也可用与第1基板和第2基板同样的材料。 The separator may also be the same material as the first substrate and the second substrate. 另外,隔板也可起到将第1基板和第2基板粘合的粘合剂的作用。 Further, the separator can also play the role of the first substrate and the second substrate bonding adhesive.

作为作用极,可使用在电子传递介质氧化时其自身不被氧化的导电材料。 As the working electrode, when the conductive material is oxidized electron transfer mediator is not oxidized itself may be used. 作为对极,只要用钯、金和白金等贵金属以及石墨等的通常所用的导电材料就可以。 As the conductive electrode material generally used, as long as the palladium, gold and platinum and other precious metals such as graphite can be. 其中,优选贵金属为主要成分来作为作用极和对极。 Wherein the noble metal as a major component is preferably used as the working electrode and the counter electrode. 这样的话,就能更精细地加工电极,由此,就可能高精度化和减少检测体的量。 In this case, the electrode can be more finely processed, whereby it is possible to reduce the amount and precision of the sample.

在本实施方式中,用光刻工艺作为电极系的制作方法,但对此没有限制。 In the present embodiment, a photolithography process using a manufacturing method of the electrode system, but there is no limitation. 例如:将贵金属溅镀到基板上形成贵金属膜,用激光修整此贵金属膜,从而就可形成电极。 For example: noble metal is formed on the substrate by sputtering a noble metal film, by laser trimming This noble metal film, whereby an electrode can be formed.

作为氧化还原酶,可以用对应于试样中所含测定对象基质的酶物质,例如:蔗糖脱氢酶、葡萄糖氧化酶、葡萄糖脱氢酶、醇氧化酶、乳酸氧化酶、胆固醇氧化酶、黄嘌呤氧化酶、氨基酸氧化酶等。 As the oxidoreductase, may be used in the sample corresponding to an enzyme target substances contained in the matrix, for example: glucose dehydrogenase, glucose oxidase, glucose dehydrogenase, alcohol oxidase, lactate oxidase, cholesterol oxidase, yellow purine oxidase, amino acid oxidase.

试剂系也可以含有亲水性高分子。 Reagent system may contain a hydrophilic polymer. 作为亲水性高分子可以用各种物质,例如:羟乙基纤维素、羟丙基纤维素、甲基纤维素、乙基纤维素、乙基羟乙基纤维素、羧甲基纤维素、聚乙烯吡咯烷酮、聚乙烯醇、聚赖氨酸等的聚氨基酸、聚苯乙烯磺酸、明胶及其衍生物、聚丙烯酸及其盐、聚甲基丙烯酸及其盐、淀粉及其衍生物、马来酸酐或是其盐的聚合物。 As hydrophilic polymer can be a variety of substances, for example: hydroxyethyl cellulose, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose, ethyl hydroxyethyl cellulose, carboxymethyl cellulose, polyvinyl pyrrolidone, polyvinyl alcohol, polyamino acid such as polylysine, polystyrene sulfonate, gelatin and derivatives thereof, polyacrylic acid and its salts, polymethacrylic acid and salts thereof, starch and derivatives thereof, MA anhydride polymer or a salt thereof. 其中优选羧甲基纤维素、羟乙基纤维素和羟丙基纤维素。 Wherein preferably carboxymethyl cellulose, hydroxyethyl cellulose and hydroxypropyl cellulose.

以下,通过实施例来更详细地说明本发明。 Hereinafter, the present invention is described in more detail by way of examples.

实施例1按照实施方式1中所示的结构制作葡萄糖传感器。 Example 1 A glucose sensor in accordance with the structure shown in Embodiment 1. 在本实施例中,作用极11和第1对极13是由65片以1μmm间隔,每片宽为5μm分支片组成的梳子状电极,作用极和对极以5μm间隔交替排列。 In the present embodiment, the working electrode 11 and the first electrode 13, working electrode and the counter electrode 65 are alternately arranged on the comb-like electrodes 1μmm intervals, each branch piece sheet width 5μm 5μm composed of at intervals.

将含有GOD及铁氰化钾水溶液滴在第1基板1的电极系上后,干燥形成试剂层,另外,在试剂层上,形成含界面活性剂卵磷脂的界面活性剂层。 The aqueous solution containing GOD and potassium ferricyanide was dropped on the electrode system after the first substrate 1 and dried to form a reagent layer, Further, in the reagent layer and surfactant layer forming surfactant-containing lecithin.

接着,把含有葡萄糖一定量的溶液作为试样,测定其中的葡萄糖浓度。 Subsequently, the amount of solution containing glucose as a sample, where the measured glucose concentration. 在本实施例中,使第1对极13和第2对极33短路来作为对极。 In the present embodiment, the electrode 13 and the second electrode 33 are short-circuited to the first electrode 1 as a pair. 将试样从试样供给口23供给到试样供给通路里,供给试样25秒钟后,以对极为基准给作用极施加300mv的电压,施加电压5秒钟后,测定作用极11和对极之间的通过电流值,通过电流/电压变换电路78将此电流值转换成电压值,此电压值就成为表示电极之间通过电流大小的指标。 A sample was supplied from the sample supply port to the sample supply passage 23, the supply of the sample after 25 seconds, the reference electrode to a voltage applied extremely 300mv to effect, voltage is applied for 5 seconds, and to determine the effect of electrode 11 the current value between the electrodes, converted by the current / voltage conversion circuit 78 this current value into a voltage value, the voltage value becomes the current through the indicator electrode between the FIG. 结果就可以观察到和试样中葡萄糖浓度成比例的电流应答。 The results can be observed and the glucose concentration in the sample is proportional to the current response.

作为比较例,仅用第1对极13来作为对极的传感器也可以进行同样的应答测定。 As a comparative example, only the first counter electrode 13 as a sensor electrode may perform the same response assays. 此情况下,关掉开关75,而开关76开着。 In this case, turn off the switch 75, the switch 76 open.

这样,就观察到了实施例1和比较例的两个传感器的和试样中的葡萄糖浓度成比例的电流应答。 Thus, the observed proportional to the concentration of glucose sensors and two samples in Example 1 and Comparative Example of the embodiment of the current response. 但是,实施例1的生物传感器可以得到比比较例的生物传感器更高的应答值。 However, embodiments of the biosensor 1 can be higher than the biosensor of Comparative Example response value. 作为高灵敏度化原因被认为是由于在实施例1中,也可以通过第2对极的作用将于作用极的垂直方向上扩散的电子传递介质的还原体在第2对极上被氧化,以及由于在作用极上抑制了扩散层生长,第2对极上的氧化还原种的浓度可以变为由传感器应答来加以反映等的缘故。 Is considered as a high sensitivity is due in Example 1, by reducing the body electronics may be diffused in the vertical direction of the second counter electrode of the working electrode will effect transfer medium on the electrode is oxidized in the second pair, and Since the effect of suppressing the diffusion electrode layer is grown on the oxide on the second counter electrode may be changed to the concentration of reduced species, because of the response to be reflected by a sensor or the like.

实施例2按照实施方式2中所示的结构制作葡萄糖传感器。 Glucose sensor prepared in Example 2 according to the configuration shown in the second embodiment. 在本实施例中,第1作用极41和第2对极63是具有32片,每片宽度为5μm,间隔为15μm的分支片的梳子状电极,第2作用极61和第1对极43是具有33片每片宽度为5μm,间隔为15μm的分支片的梳子状电极。 In the present embodiment, the first working electrode 41 and the second counter electrode 63 is with 32 tablets, each width of 5 m, the interval of comb-shaped electrode branches sheet 15μm, and the second working electrode 61 and the first counter electrode 43 is 5μm, 15μm sheet of spacer is a branched comb-shaped electrode 33 having a width of each piece. 第1作用极以5μm间隔和第1对极交替排列,而第2作用极和第2对极也是以5μm间隔交替排列。 Poles are alternately arranged at an extremely 5μm 5μm spacing interval and the first counter electrode are arranged alternately, and the second working electrode 1 and the second to the first action. 由此,第1作用极和第2对极,还有第2作用极和第1对极,都各自向对配置。 Thereby, the first working electrode and second counter electrode, and the second working electrode and a counter electrode, both to each configuration. 试剂层和界面活性剂层的构成与实施例1相同。 The same configuration as in Example 1 the reagent layer and surfactant layer.

如同实施例1,把含有一定量葡萄糖的溶液作为试样,测定其中的葡萄糖浓度。 As in Example 1, the solution contains a certain amount of glucose as a sample, in which the measured glucose concentration. 在本实施例中,使第1对极43和第2对极63短路形成对极,将第2作用极41和第1作用极61短路来形成作用极。 In the present embodiment, the counter electrode 43 and the second electrode 63 to short-circuit the first counter electrode is formed, the second working electrode 41 and the electrode 61 short-circuiting the first action to form a working electrode. 从试样供给口52将试样供给到试样供给通路内,25秒钟后,以对极为基准,施加300mv电压于作用极上,结果是,可以得到比实施例1中使用比较例传感器更高的应答值。 A sample from the sample supply port 52 is supplied to the sample supply pathway, after 25 seconds, in order to extremely reference voltage is 300mv is applied to the working electrode, a result can be obtained than that used in Example 1 Comparative Example sensor is more high response value.

实施例3除了如图6所示添加参比极15以外,其余的均与实施例1一样制作传感器。 Example 3 except for the addition than the reference electrode 15 shown in FIG. 6, the rest were the same as in Example 1 produced sensor. 按照图9所示将传感器安装在测定装置上,从试样供给口23将到试样供给试样供给通路内,试样供给25秒钟后,以参比极15为基准施加300mv电压于作用极11上,施加电压5秒钟后,测定作用极11和对极之间的通过电流值,通过电流/电压变换电路78将此电流值变换为电压值。 Mounting the sensor in FIG. 9 as shown in the measuring device, from the sample supply port 23 to supply a sample into the sample supply passage 25 seconds after sample supply to the reference electrode 15 at a voltage of 300mv effect Jizhunshijia upper electrode 11, voltage is applied for 5 seconds, measured by the working electrode 11 and the counter electrode between the current value, a current value of 78 by this current / voltage conversion circuit is converted into a voltage value.

实施例3传感器和实施例1中的传感器一样能提供高灵敏度的应答,另外,因为有了参比极,和二电极方式相比就能够稳定作用极的电位,由此,就降低了应答值的偏离。 Example 3 Example 1 of the sensor and sensor embodiment can provide the same response with high sensitivity, further, because the reference electrode, and the electrode system than two can be compared to a stable potential of the working electrode, thereby, reduces the response value deviation.

在上述实施例中,作用极和对极的每片分支片宽度为10μm,而同一基板上的作用极和对极的距离为5μm,但是,对此没有限制。 In the above embodiment, the working electrode and the counter electrode branches per tablet slice width is 10 m, and the working electrode on the same substrate and the electrode distance is 5 m, however, there is no limitation. 另外,从试样的供给到电压的施加要有25秒钟,对此也没有限制,酶反应进行时间要求有能得到和试样中基质浓度相关的电流应答的程度就行,最好在180秒钟以下。 Further, the sample is applied to the voltage supplied from a least 25 seconds, there is no restriction to this, the enzyme reaction time required to get the degree of the response current and substrate concentration in the sample related to the line, preferably 180 seconds bell less.

对电极系施加300mv的电压,但对此无限制,只要有使电子传递介质在作用极上进行电极反应的电压就行。 300mv voltage is applied to the electrode system, but this limitation, as long as the electron transfer mediator in the electrode reaction on the working electrode voltage line.

对于作用极和对极的距离,形成在同一基板上的作用极分支片和对极分支片之间的距离最好在1-50μm的范围:第1基板的电极和第2基板的电极之间的距离由隔板的厚度来决定的,隔板的厚度最好在1-50μm的范围。 For the working electrode and the effect electrode distance, is formed on the same substrate the pole pieces branch of 1-50μm and preferably in the range of a distance between the pole pieces branches: between the electrode and the second substrate a first substrate , by a distance in the range of thickness of the separator to determine the thickness of the separator is preferably 1-50μm.

在实施例中,用铁氰化钾来作为电子传递介质,但对此没有限制,也可以用对苯醌、吩嗪甲基硫酸盐、亚甲蓝、二茂铁衍生物等。 In an embodiment, potassium ferricyanide as an electron transfer mediator, but there is no limitation, may be a p-benzoquinone, phenazine methosulfate, methylene blue, ferrocene derivatives. 另外,在氧作为电子传递介质的情况下,也能得到电流应答。 Further, in the case of oxygen as an electron transfer mediator, current response can be obtained. 也可以使用上述的两种以上来作为电子传递介质,上述实施例中,使第1对极和第2对极短路以形成对极,但对此无限制,也可以使第1对极和第2对极独立运作。 It may also be used as the above-mentioned two or more electron transfer mediator, the above-described embodiment, the first counter electrode and the second electrode of the counter electrode to form a short circuit, but this limitation may be on the first electrode and the second two pairs of poles to operate independently. 例如:给第1对极施加可使电子传递介质还原的定位,而即使只使用第2对极来作为对极也是可行的。 For example: a first counter electrode is applied to the electron transfer mediator can restore location, and even if only the second counter electrode as a counter electrode is also possible.

上述实施例中,用了β-D-葡萄糖水溶液作为试样,但对此没有限制。 The above-described embodiment, with the β-D- aqueous glucose solution as a sample, but there is no limitation. 例如,也可以采用全血、血浆、血清、间质液、唾液和尿等的生的体试样。 For example, whole blood, plasma, serum, interstitial fluid, saliva and urine and other body samples may be used raw. 试样为全血的情况是,例如,通过穿刺指尖和腕部皮肤而取得的毛细血或是静脉血、动脉血等。 Whole blood sample is the case, for example, capillary blood obtained by pricking the finger and the wrist skin or blood, arterial blood and the like.

产业上应用的可能性通过上述本发明,可以制得即使是极微的试样量,也可以得到良好应答的高灵敏度生物传感器。 INDUSTRIAL APPLICABILITY The present invention described above can be prepared even minimal amounts of sample, a high sensitivity can be obtained a favorable response of the biosensor.

Claims (4)

1.一种生物传感器,它包括具有分支成多个的作用极和分支成多个的第1对极、将各分支片交替排列的第1绝缘性基板,具有第2对极、配置于和第1绝缘性基板相对位置上的第2绝缘性基板,含有氧化还原酶和电子传递介质的试剂系以及形成在第1和第2绝缘性基板之间的试样供给通路,在供给通路内交替排列的作用极和第1对极的分支片、第2对极和试剂系外露。 A biosensor, comprising a first counter electrode, the first working electrode and the insulating substrate having a plurality of branches branched into a plurality of sheets are alternately arranged in the respective branch, having a second counter electrode disposed on and the first insulating substrate opposite the second insulating substrate at a position, based reagent containing an oxidoreductase and an electron transfer mediator and a sample supply pathway formed between the first and second insulating substrates, alternately in the supply passage the arrangement of the first working electrode and the counter electrode branch piece, exposed to the second electrode and reagent system.
2.根据权利要求1所述的生物传感器,其特征在于,在所述试样供给通路内,第2对极只配置在与作用极相对的位置上。 2. The biosensor according to claim 1, wherein, in said sample supply passage, only the second counter electrode disposed at a position opposite to the working electrode.
3.一种生物传感器,它包括具有分支成多个的第1作用极和分支成多个的第1对极、将各分支片交替排列的第1绝缘性基板,具有分支成多个的第2作用极和分支成多个的第2对极、将各分支片交替排列的第2绝缘性基板,含有氧化还原酶和电子传递介质的试剂系以及形成在第1和第2绝缘基板之间的试样供给通路,在试样供给通路内交替排列的第1作用极和第1对极的分支片、交替排列的第2作用极和第2对极的分支片以及试剂系外露。 A biosensor which includes a working electrode is branched into a first branch and a plurality of a first plurality of counter electrode, the first insulating substrate 1 are alternately arranged in the respective branch piece, having a plurality of first branches 2 is branched into a plurality of working electrode and second counter electrode, the second insulating substrate pieces alternately arranged in each branch, the reagent system comprising an oxidoreductase and an electron transfer medium, and formed between the first and second insulating substrate the sample supply passage, the supply passage in the sample are alternately arranged in a first working electrode and the counter electrode of the first branch piece, the role of the second electrode are alternately arranged and exposed on the second electrode sheet, and branch-based reagents.
4.根据权利要求3所述的生物传感器,其特征在于,第2对极配置在和第1作用极相对的位置上,且第2作用极配置在和第1对极相对的位置上。 4. The biosensor according to claim 3, wherein the second position and a counter electrode disposed opposing action on the first electrode and the second working electrode disposed at a position opposite to the first and the counter electrode.
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